JP5685542B2 - Water-insoluble polymer for colon targeting: starch-based film coating - Google Patents

Description

  The present invention relates to formulations for the controlled delivery of active ingredient (s). The invention also relates to its use and method of manufacture.

  Colon targeting can be very useful for many drug therapies, including the treatment of inflammatory bowel diseases such as Crohn's disease (CD) and ulcerative colitis (UC).

  When topically acting drugs are administered orally by conventional pharmaceutical formulations, the latter is rapidly dissolved in the stomach contents and the drug is released and possibly absorbed into the bloodstream. This increases the systemic drug concentration, which increases the risk of undesirable side effects and at the same time decreases the drug concentration at the site of action in the colon, resulting in poor therapeutic efficiency. These limitations can be overcome if drug release is suppressed in the stomach and small intestine and time controlled in the colon. This type of site-specific drug delivery to the colon may also provide an interesting opportunity for protein and peptide drugs to be absorbed into the systemic bloodstream by oral administration.

  To enable colon targeting, for example, the drug can be embedded in a polymeric matrix former, or a drug-carrying tablet or pellet, such as a sphere with a diameter of about 0.5-1 mm It can be a bead or it can be coated with a polymer film. In the upper gastrointestinal tract (GIT), the permeability of the polymer network to the drug must be low, but the macromolecular barrier must become permeable when it reaches the colon. This increase in drug permeability of the polymer network at the site of action may be (i) a change in the pH of the GIT contents, (ii) a change in the quality and / or quantity of the enzyme along the GIT, or (iii) a given Can be induced by significant structural changes in the formulation that occur after a lag time (e.g., crack formation of a low permeability film coating that provides a pulsatile drug release pattern). '' 'As another option, drug release is already initiated in the stomach and persists throughout the GIT at a rate low enough to ensure that the drug is still in the formulation when it reaches the colon. You may be made to do.

  An attempt to solve the problem of colon targeting is disclosed in US Patent Application Publication No. 2005/0220861 relating to a controlled release formulation for delivering prednisolone sodium metasulfobenzoate. The formulation comprises prednisolone sodium metasulfobenzoate surrounded by a coating comprising glassy amylose, ethyl cellulose and dibutyl sebacate. However, the ratio of amylose to ethyl cellulose is (1: 3.5) to (1: 4.5), and amylose is corn or maize amylose. In contrast to US 2005/0220861, the system according to the present invention is adapted to the disease state of a patient. This is a very important aspect. This is because the formulation must become more permeable to the drug when it reaches the colon in order to allow colon targeting. This can be ensured, for example, by preferentially degrading compounds that interfere with rapid drug release in the upper gastrointestinal tract. This site-specific degradation can rely on significant quality and quantity differences between enzymes present in the upper gastrointestinal tract and those in the colon. The compound must not be degraded in the upper gastrointestinal tract (and must interfere with drug release), but it must be degraded in the colon (and thus allow drug release). The performance of this type of advanced drug delivery system is basically dependent on the environmental conditions in the patient's colon, and in particular on the type and concentration of enzymes present in the colon. It is well known and well documented that disease states can significantly affect the quality and quantity of enzyme-secreting microbiota in the gastrointestinal tract. This is especially true in the case of microbiota in the colon of patients suffering from inflammatory bowel disease. That is, for example, the quality and quantity of enzymes present in the patient's colon can be significantly different from that of healthy subjects. Thus, the performance of this type of drug delivery system can be significantly affected by the disease state. Systems that rely on preferential degradation by enzymes that are not present in sufficient concentrations in disease states within the patient's colon do not work well. The present invention reports for the first time a formulation that allows controlled delivery of the active ingredient under pathophysiological conditions, ie in the stool of a patient suffering from inflammatory bowel disease. Thus, the performance of this formulation is guaranteed in vivo under a given pathophysiological condition. This is critical for the response and safety of the treatment.

  U.S. Pat. No. 6,534,549 describes a substantially water-insoluble film-forming polymer in a solvent system comprising (1) water and (2) a water-miscible organic solvent capable of dissolving the film-forming polymer alone. The present invention relates to a process for producing a controlled release composition comprising contacting a mixture of amylose and amylose with an active material and drying the resulting composition. This formulation is particularly suitable for delivering therapeutic agents to the colon. In contrast to the present invention, the disclosure deals with drug delivery systems prepared using organic solvents. This is not the case with the present invention. The use of organic solvents has several problems including toxicity and environmental problems as well as explosion hazard. Furthermore, the use of amylose involves the extraction of this polymer and its stabilization. Amylose is extracted from the starch after the hydrolysis and purification steps. This process is complex and difficult to use at the industrial level. This composition does not take into account the rate of drug release in patients suffering from inflammatory bowel disease. It should be pointed out that the type and amount of bacteria present in the colon of patients with inflammatory bowel disease can differ significantly from those of healthy subjects. Thus, the type and amount of enzymes that are secreted by these bacteria and contact the drug delivery system can be significantly different. As a result, the performance of the drug delivery system can be significantly different.

US Patent Application Publication No. 2005/0220861 US Pat. No. 6,534,549

“Composition, structure, function and chemical modification of legume starch: a review”. Hoover et al. "Development of New Pea Starch" CL-Heydley et al. (Proceedings of the Symposium of the Industrial Biochemistry and Biotechnology Group of the Biochemical Soc. 77).

  The object of the present invention is an active ingredient, such as, but not limited to, an active pharmaceutical ingredient, biologically active ingredient, chemically active ingredient, vegetative active ingredient, agricultural active ingredient, or nutritionally active ingredient It is to provide a delivery formulation that controls the rate and extent of delivery.

  Another object of the present invention is to provide a new polymeric film coating that allows site-specific drug targeting to the colon and can be used not only in patients with healthy colon but also in patients with inflammatory bowel disease. Is to provide.

  A further object of the present invention occurs in the formulation due to water permeation into the system sufficient to withstand shear stresses (due to gastrointestinal motility) experienced in the upper GIT and upon contact with aqueous media. It is to provide a new polymeric film coating with mechanical stability sufficient to withstand significant hydrostatic pressure that may be Indeed, in known polymer coatings, the problem of accidental crack formation can lead to premature drug release through water-filled channels.

  It is a further object of the present invention to provide new polymeric film coatings that can be tailored to the specific needs of specific types of drug treatments, such as drug osmotic activity and dosage.

The present invention
o at least a water-insoluble polymer; o starch having an amylose content of 20-45%, preferably 25-44%, even more preferably 30-40%, a modified starch having an amylose content of 50-80%, and legumes Provided is a controlled release delivery formulation for controlled release of an active ingredient comprising an active ingredient coated with a polymer mixture with a starch composition comprising at least one ingredient selected from the group consisting of plant starch.

  Preferably, the controlled release formulation is an oral formulation and has gastric tolerance. In preferred embodiments, the controlled release pharmaceutical formulation is in solid, liquid, or semi-liquid form. Advantageously, the controlled release pharmaceutical formulation is a solid dispersion. According to the present invention, the polymer mixture is a homogeneous mixture of a water-insoluble polymer and a starch composition, said starch does not form microparticles in the water-insoluble polymer.

  In one embodiment of the invention, the polymer mixture of the controlled release delivery formulation is a coating mixture, the controlled release delivery formulation comprises a core and the active ingredient is dispersed or dispersed in the core and / or in the coating mixture. Dissolved.

  In a further embodiment of the invention, the starch composition: water insoluble polymer ratio of the controlled release delivery formulation is 1: 2 to 1: 8, preferably 1: 3 to 1: 6, more preferably 1: 4 to 1. : 5.

  Preferably, the starch composition has a starch content of at least 50%, preferably 70-100%, more preferably 70-100%, and even more preferably 90-100%.

  Typically, the starch composition exhibits an amylose content of 20-45%, preferably 25-44%, more preferably 32-40%, and this percentage is present in the composition. Expressed as a dry weight based on the dry weight of the starch.

  In a further embodiment of the invention, the starch composition comprises at least one legume starch or cereal starch.

  Preferably, the leguminous plant is selected from the group consisting of peas, green beans, broad beans (broad beans and horse beans).

  According to another advantageous alternative form, the legumes are plants that give seeds containing at least 25% by weight of starch (preferably dry / dry), for example various peas or broad beans. It is. Preferably, the legume starch is a granular legume starch.

  Advantageously, the legume is a pea. Pea starch granules have two attributes. The first attribute is a large granule diameter (eg larger than corn starch granules) that results in improved granule surface area and thus improved contact with water and microflora enzymes in the colon. In addition, pea starch granules have a high swelling capacity that results in an improvement of their surface area and thus an improved digestibility of the granules and consequently an improved release of the active ingredient in the colon.

  According to another advantageous option, the legume starch is a natural legume starch.

  Advantageously, this starch content of the starch composition is greater than 90% (dry / dry). Specifically, it may be more than 95%, preferably more than 98%.

  According to the present invention, the modified starch is preferably stabilized. Indeed, according to a preferred embodiment of the present invention, a particularly suitable chemical treatment for the preparation of a film-forming composition is a “stabilization” treatment. Normal stabilizing modification can be achieved by esterifying or etherifying some of the hydroxyl groups along the starch chain. Preferably, the modified starch is hydroxypropylated and / or acetylated, and optionally a fluidization treatment, which is a chemical hydrolysis treatment or an enzymatic hydrolysis treatment, can be added to these treatments. is there. Preferably, the modified starch is fluidized by, for example, acid treatment. Thus, the starch composition according to the invention advantageously comprises at least one stabilized starch, preferably a hydroxypropylated starch exhibiting a degree of substitution (DS) of at most 0.2. The term “DS” is considered in the present invention to mean the average number of hydroxypropyl groups per 10 anhydroglucose units. This average number is determined by standard analytical methods well known to those skilled in the art.

  In a further embodiment of the invention, the starch composition comprises xylo-oligosaccharides, inulin, oligofructose, fructooligosaccharide (FOS), lactulose, galactomannan and suitable hydrolysates thereof, indigestible polydextrose, indigestible dextrin and Its partial hydrolysates, trans-galacto-oligosaccharides (GOS), xylo-oligosaccharides (XOS), acemannan, lentinan or β-glucan and their partial hydrolysates, polysaccharide-K (PSK), and indigestible maltodextrins and The partial hydrolyzate may preferably additionally comprise at least one indigestible polysaccharide selected from the group consisting of indigestible dextrin or indigestible maltodextrin.

  According to the present invention, indigestible maltodextrin or indigestible dextrin is 15-35% 1-> 6 glucoside bond, less than 20% reducing sugar content, less than 5 multimolecular index, and at most 4500 g. Number average molecular mass Mn equal to / mol.

  According to one variant, all or part of the indigestible maltodextrin is hydrogenated.

  According to one variant, the core has a coating level of 5% to 30%, preferably 10% to 20%.

  In a further embodiment, the polymer mixture includes a plasticizer. Preferably, the plasticizer content is 25% to 30% w / w based on the water-insoluble polymer content.

  Preferably, the water-insoluble polymer is ethyl cellulose, cellulose derivative, acrylic acid and / or methacrylic ester polymer, acrylate or methacrylate polymer or copolymer, polyvinyl ester, starch derivative, polyvinyl acetate, polyacrylic ester, butadiene styrene copolymer, methacrylic ester, It is selected from the group consisting of acid ester copolymer, cellulose acetate phthalate, polyvinyl acetate phthalate, shellac, methacrylic acid copolymer, cellulose acetate trimellitate, hydroxypropylmethylcellulose phthalate, zein, starch acetate.

  According to a further embodiment, the plasticizer is a water soluble plasticizer. Preferably, the water-soluble plasticizer is used alone or as a mixture with each other, polyol (glycerin, propylene glycol, polyethylene glycol), organic ester (phthalate ester, dibutyl sebacate, citrate ester, triacetin), oil / glyceride ( Castor oil, acetylated monoglyceride, fractionated coconut oil) and soy lecithin.

  In a preferred embodiment, the controlled release formulation is a multiparticulate formulation.

The present invention also provides controlled release delivery for controlled release of an active ingredient in the colon of a patient having an imbalance of intracolonic microbiota or in the colon of a healthy subject as claimed in claims 1-12. A method for producing a formulation is provided. However, the method
o The following substances:
At least one water-insoluble polymer; starch having an amylose content of 20-45%, preferably 25-44%, even more preferably 30-40%, modified starch, amylose content of 50-80% Forming a polymer mixture with a starch composition comprising a modified starch having and at least one component selected from the group consisting of legume starches;
o coating the active ingredient with a polymer mixture;
including.

  In a further embodiment, the step of coating the active ingredient is a step of coating the core with the active ingredient dispersed or dissolved in the core and / or the step of coating the active ingredient is in the polymer mixture. This is a step of dispersing or dissolving the active ingredient.

  The condition in the gastrointestinal tract of patients suffering from inflammatory bowel disease (eg Crohn's disease and ulcerative colitis) can be significantly different from that of healthy subjects. Intra-individual and inter-individual variability can be quite large with respect to the pH of the GIT content, the type and concentration of enzyme-secreting bacteria, and the transit time within the various GIT segments. For example, in general, there is a significant amount of bifidobacteria in the colon of a healthy subject, and it is possible to degrade complex polysaccharides with multiple extracellular glycosidases. However, in disease states, their concentrations can be significantly reduced. For example, fecal glycosidase activity (specifically β-D-galactosidase activity) is reduced in patients with Crohn's disease, and the metabolic activity of the colonic microbiota is strongly disturbed in active disease states It was shown that. Thus, the effects of pathophysiology can be significant and can lead to drug treatment failures.

  In order to avoid treatment failure for patients suffering from inflammatory bowel disease, site-specific drug delivery systems must be adapted to a given condition of the patient's colon. For example, polymeric film coatings that are degraded by enzymes present in sufficient amounts in the feces of Crohn's disease patients and ulcerative colitis patients may be used. However, it is still unclear which type of polymer meets these prerequisites.

Water content of thin films composed of various types of polymer blends (shown in the figure) upon exposure to (a) 0.1 M HCl and (b) phosphate buffer pH 6.8. A film composed solely of plasticized ethylcellulose is shown for comparison. Dry mass of thin films composed of various types of polymer blends (shown in the figure) upon exposure to (a) 0.1 M HCl and (b) phosphate buffer pH 6.8. A film composed solely of plasticized ethylcellulose is shown for comparison. Water content of thin films composed of branched maltodextrin or pea starch blended with ethylcellulose upon exposure to phosphate buffer pH 6.8 with or without pancreatin or rat intestinal extract And dry mass. Various blended with ethylcellulose upon exposure to culture medium, culture medium inoculated with feces of healthy subjects, and culture medium inoculated with feces of Crohn's disease (CD) and ulcerative colitis (UC) patients Water content and dry mass of thin films composed of types of polysaccharides (as shown in the figure). A film composed solely of plasticized ethylcellulose is shown for comparison. Photographs of microflora generated when thin polymer films of various compositions (shown in the figure) were incubated with stool samples from patients with inflammatory bowel disease. Water uptake and dry mass loss of thin films composed of PS HP-PG: ethylcellulose blends upon exposure to 0.1 M HCl and phosphate buffer pH 6.8, respectively. The polymer blend ratio is shown in the figure. For comparison, the behavior of pure (plasticized) ethylcellulose film is also shown. Change in breaking energy of thin PS HP-PG: ethylcellulose films upon exposure to (a) 0.1 M HCl and (b) phosphate buffer pH 6.8. The polymer blend ratio is shown in the figure. For comparison, the results obtained with pure (plasticized) ethylcellulose film are also shown. Water uptake and dry mass loss of thin films composed of EURYLON® 7A-PG: ethylcellulose blends upon exposure to 0.1 M HCl and phosphate buffer pH 6.8, respectively. The polymer blend ratio is shown in the figure. For comparison, the behavior of pure (plasticized) ethylcellulose film is also shown. Change in breaking energy of thin EURYLON® 7A-PG: ethylcellulose film upon exposure to (a) 0.1 M HCl and (b) phosphate buffer pH 6.8. The polymer blend ratio is shown in the figure. For comparison, the results obtained with pure (plasticized) ethylcellulose film are also shown. Water uptake and dry mass loss of thin films composed of EURYLON® 6A-PG: ethylcellulose blends upon exposure to 0.1M HCl and phosphate buffer pH 6.8, respectively. The polymer blend ratio is shown in the figure. For comparison, the behavior of pure (plasticized) ethylcellulose film is also shown. Change in breaking energy of thin EURYLON® 6A-PG: ethylcellulose film upon exposure to (a) 0.1 M HCl and (b) phosphate buffer pH 6.8. The polymer blend ratio is shown in the figure. For comparison, the results obtained with pure (plasticized) ethylcellulose film are also shown. Water uptake and dry mass loss of thin films composed of EULYLON® 6 HP-PG: ethylcellulose blends upon exposure to 0.1 M HCl and phosphate buffer pH 6.8, respectively. The polymer blend ratio is shown in the figure. For comparison, the behavior of pure (plasticized) ethylcellulose film is also shown. Change in breaking energy of thin EURYLON® 6 HP-PG: ethylcellulose film upon exposure to (a) 0.1 M HCl and (b) phosphate buffer pH 6.8. The polymer blend ratio is shown in the figure. For comparison, the results obtained with pure (plasticized) ethylcellulose film are also shown. Water uptake rate of thin pea starch: ethylcellulose film upon exposure to (a) 0.1 M HCl and (b) phosphate buffer pH 6.8. The polymer: polymer blend ratio (w: w) is shown in the figure. Dry mass loss rate of thin pea starch: ethylcellulose films upon exposure to (a) 0.1 M HCl and (b) phosphate buffer pH 6.8. The polymer: polymer blend ratio (w: w) is shown in the figure. Mechanical properties of dry, thin pea starch: ethylcellulose films: (a) puncture strength at break, (b)% elongation at break, and (c) energy at break. The polymer: polymer blend ratio (w: w) is shown on the x-axis. Change in break energy of thin pea starch: ethylcellulose films upon exposure to (a) 0.1 M HCl (2 hours) or (b) phosphate buffer pH 6.8 (8 hours) at 37 ° C. The polymer: polymer blend ratio (w: w) is shown in the figure. Pea starch under conditions simulating passage through the upper gastrointestinal tract, i.e., 2 hours exposure to 0.1 M HCl followed by 9 hours exposure to phosphate buffer pH 6.8 : Drug release from pellets coated with ethylcellulose 1: 2. The coating level, as well as the absence (solid curve) and presence (dotted curve) of the enzyme (0.32% pepsin at low pH, 1% pancreatin at neutral pH) are shown in the figure. Under conditions simulating passage through the upper gastrointestinal tract, i.e., 2 hours exposure to 0.1 M HCl followed by 9 hours exposure to phosphate buffer pH 6.8. Effect of pea starch: ethylcellulose blend ratio (w: w) and coating level (shown in the figure) on drug release from coated pellets. The solid / dotted curve represents the absence / presence of the enzyme (0.32% pepsin at low pH, 1% pancreatin at neutral pH). Under conditions simulating passage through the entire gastrointestinal tract, ie, 2 hours exposure to 0.1 M HCl followed by 9 hours exposure to phosphate buffer pH 6.8 (a 15% or 20% (as specified) after 10 hours of exposure to culture medium inoculated with a new stool sample from a patient with inflammatory bowel disease or (b) culture medium inoculated with bifidobacteria Release from pellets coated with pea starch: ethylcellulose 1: 4 at the following coating levels: For comparison, drug release upon exposure to culture media without stool samples is also shown in (a) (dotted curve). Storage stability of pellets coated with 10%, 15% and 20% pea starch: ethylcellulose 1: 4: 0.1 M before (solid curve) and after (solid curve) after 1 year of open storage Drug release in HCl (2 hours) and in phosphate buffer pH 6.8 (9 hours). (A) Medium only in rectum (negative control group), (B) TNBS in rectum (positive control group), (C) TNBS in rectum and orally BMD: EC coated pellet, or (D) Macroscopic appearance of the colon of a rat administered TNBS intrarectally and orally Pentasa® pellet. Only TNBS / medium was administered intrarectally on day 3. The oral dose of 5-ASA was 150 mg / kg / day. (Ii) TNBS in the rectum, (ii) TNBS in the rectum and orally Asacol® pellets, (iii) TNBS in the rectum and orally Pentasa® pellets, (iv) rectum Ameho score of rats administered with TNBS and oral pea starch: ethylcellulose coated pellets, (v) TNBS and oral BMD: ethylcellulose coated pellets rectally. TNBS was administered intrarectally on day 3. The oral dose of 5-ASA was 150 mg / kg / day. Representative histological section of rat colon tissue (normal transmural section, × 200). Various layers are shown: M-mucosa, SM-submucosa, Mu-muscle layer.

  Table 1: Bacterial concentration [log CFU / g] in stool samples studied in healthy subjects and patients with inflammatory bowel disease.

  Table 2: Effect of the type of polysaccharide blended with ethylcellulose and the blend ratio of polysaccharide: ethylcellulose on the mechanical properties of thin films in the dry state at room temperature.

  Table 3: Dissolution media used to simulate pH grading along GIT.

DETAILED DESCRIPTION OF THE INVENTION In describing and claiming the present invention, the following terminology will be used in accordance with the definitions set out herein.

  As used herein, the term “active ingredient”, “drug”, or “pharmacologically active ingredient” or any other similar term refers to (1) an organism that has a prophylactic effect on the organism and is undesirable Prevent inflammatory effects, eg prevent infection, (2) alleviate the pathology caused by the disease, eg alleviate pain or inflammation caused as a result of the disease, and / or (3 ) Inducing a desired biological or pharmacological effect that may include, but is not limited to, mitigating, reducing, or completely eliminating disease from an organism Any chemical or biological material or compound suitable for administration by methods already known in the art and / or by the methods taught in the present invention. Taste. The effect may be local, such as providing a local anesthetic effect, or systemic.

  As used herein, the expression “imbalance of microbiota in the colon”, also referred to as enterotoxemia or intestinal bacterial overgrowth, is used in the present invention to refer to qualitative or quantitative microorganisms in the gastrointestinal tract. It means imbalance. This phenomenon is reflected in the quality and quantity of enzymes present in the colon. Specifically, this altered microflora is observed in the colon of patients suffering from inflammatory bowel diseases such as Crohn's disease (CD) and ulcerative colitis (UC).

  As used herein, the term “controlled release delivery” or “controlled release” means that the release of the active ingredient from the formulation is controlled with respect to time or with respect to the site of delivery.

  The expression “modified starch” is to be interpreted broadly, and this expression means, for example, reticulated starch or acetylated starch or hydroxypropylated starch.

  The term “coating” is used herein to encompass capsules surrounding fluids and / or solids in addition to coatings on solid carriers, and the term “coated” is used as well. It is done.

  The expression “water-insoluble polymer” is to be interpreted broadly and refers to polymers that are not completely soluble in water, such as ethyl cellulose, certain starch derivatives, or acrylic / methacrylic acid derivatives. means.

  The term “indigestible polysaccharide” as used in the present invention means that it is not digested or partially digested in the intestine by the action of acids or digestive enzymes present in the human upper digestive tract (small intestine and stomach). However, it means a saccharide that is at least partially fermented by the human gut microbiota. Indigestible water-soluble polysaccharides that can be used in preferred embodiments of the present invention include xylo-oligosaccharides, inulin, oligofructose, fructo-oligosaccharides (FOS), lactulose, galactomannan and suitable hydrolysates thereof, indigestible polydextrose. Indigestible dextrin and its partial hydrolysates, trans-galacto-oligosaccharides (GOS), xylo-oligosaccharides (XOS), acemannan, lentinan or β-glucan and their partial hydrolysates, polysaccharide-K (PSK), and Indigestible maltodextrin and its partial hydrolyzate.

  Polysaccharide-K is also known as polysaccharide-krestin (PSK) in Japan and polysaccharide-peptide (PS-P) in China. Both have the same chemical and structural properties. PSK is a proteoglycan found in the porous fungus Trametes versicolor, containing about 35% carbohydrate (91% β-glucan), 35% protein, the rest being sugar, amino acids, And free residue like moisture. PSK is a mixture of polysaccharides covalently linked to various peptides and has an average molecular weight of 100 kilodaltons. The polysaccharide component belongs to β-glucans composed of glucopyranose units. As a result of structural analysis, PSK has 1,4-glucan configuration having a branch at positions 3 and 6 as a main glucoside moiety with a frequency of one branch per several residues of 1 to 4 bonds. It was shown that.

  As used herein, the term “cereal” as used herein refers to any plant belonging to the Gramineae, preferably wheat, rice, rye, oats, barley, corn, sorghum, and millet. It shall mean a thing.

  The term “leguminous plant” is used in the present invention to refer to any plant belonging to the family Caesalpinaceae, Mimosaceae, or Papillionaceae, in particular any belonging to the Pirionaceae family. Plant, for example, pea, green beans, broad bean, lentil, alfalfa, clover, or lupine.

  The expression “starch derivative” means starch that has been enzymatically or chemically treated.

  “Coating level” means the weight difference between an uncoated core and a coated core, which is a weight gain in percent.

  This definition is specifically described in R.D., entitled “Composition, structure, function and chemical modification of legume starch: a review”. Includes all plants listed in any one of the tables present in the paper of Hoover et al.

The term “pea” is considered to have its broadest meaning, specifically,
• all wild species of smooth peas, and • all mutants of smooth peas and wrinkle peas (but this is generally intended for the intended use of the varieties (human food, animal nutrition food, and / or other No matter what)
Is included.

  The mutant species is specifically described in a paper by C-L Heydley et al. Entitled “Development of New Pea Starch” (Proceedings of the Symposium of the Industrial Biochemistry and Biotechnology Group of Biotechnology, Biotechnology of Biotechnology, Biotechnology, Biotechnology, Biotechnology of Biotechnology, Biotechnology of Biotechnology, Biotechnology, et al. -87) as "mutant r", "mutant rb", "mutant rug3", "mutant rug4", "mutant rug5", and "mutant lam". Is referred to as.

  The term “legume starch” is extracted from a legume as defined above (however, regardless of method) and is greater than 40%, preferably greater than 50%, even more preferably 75%. It is taken to mean any composition having an excess starch content, where these percentages are expressed in dry weight based on the dry weight of the composition.

  Furthermore, it is possible to use starch that naturally exhibits an amylose content within the range selected according to the invention. In particular, starches derived from legumes may be suitable. According to the present invention, the legume starch exhibits an amylose content of less than 45%, more particularly 2020-45%, preferably 25-44%, even more preferably 32-40%.

  For the purposes of the present invention, the term “ingestible maltodextrin” refers to an indigestible glucoside bond that gives maltodextrin an additional property equivalent to dietary fiber such as “branched maltodextrin”. Means maltodextrin contained. As used herein, the term “branched maltodextrins” refers to ingestible maltodextrins as described in EP 1006128 (whose company is the owner). It shall be.

  According to a preferred variant, the branched maltodextrins have a reducing sugar content of 2% to 5% and a number average molecular mass Mn of 2000 to 3000 g / mol.

  The branched maltodextrin is obtained from AOAC Method No. When determined according to 2001-03 (2001), it has a total fiber content of 50% or more on a dry basis.

  The present invention provides a novel polymeric film coating for colon targeting that is adapted to the disease state of a patient suffering from inflammatory bowel disease.

  The novel polymer film according to the present invention functions as a substrate for colonic bacteria in both healthy patients and patients suffering from inflammatory bowel disease and will probably have a beneficial effect on the patient's GIT ecosystem. . The polymer film is specifically adapted to the condition of the target site, even in disease states, and can deliver pharmacologically active ingredients specifically to the colon.

  The invention is illustrated below with the aid of the following examples and drawings.

Example 1
A. Materials and Methods 1. Materials Branched maltodextrin (BMD) [Non-digestible glycosidic bonds: branched maltodextrins having α-1,2 and α-1,3, NUTRIOSE® FB06 Roquette Freres], pea starch (granular pea starch N-735) (35% amylose), pregelatinized hydroxypropyl pea starch (PS HP-PG) (LYCOAT® RS780), maltodextrin (MD) (GLUCIDEX® 1, Roquette Freres), EURYLON ( 7A-PG (acetylated and pregelatinized high amylose maize starch (70% amylose) (Roquette Freres, Lestrem, France), EURYLON® 6A PG (acetylated and pregelatinized high amylose maize starch) (60% amylose) (Roquette Freres, Lestrem, France) and EURYLON® 6HP-PG (hydroxypropylated and pregelatinized high amylose maize starch (60% amylose) ) (Roquette Freres, Restrem, France), aqueous ethylcellulose dispersion (Aquacoat® ECD30, FMC Biopolymer, Philadelphia, USA), triethyl citrate (TEC; Morflex®, CreansA, US) Mammalian pancreas origin = mixture containing amylase, protease and lipase, Fisher Bio block, Illkirch, France), rat intestinal extracts (rat intestinal powder containing amylase, sucrase, isomaltase, and glucosidase, Sigma-Aldrich, Isled'Abeau Chesnes, France), Columbia blood agar, beef cattle and yeast Extracts from, as well as tryptone (= casein pancreatic juice digest) (Becton Dickinson, Sparks, USA), L-cysteine hydrochloride hydrate (Acros Organics, Geel, Belgium), McConkey agar (BioMerieux, Balme-les-) Grottes, France), cysteine-added Ringer's solution (Merck, Darmstadt, Germany).

A. 2. Film Preparation Thin polymer films were prepared by casting blends of various types of aqueous polysaccharides and aqueous ethylcellulose dispersions into a Teflon mold followed by drying at 60 ° C. for 1 day. . Water soluble polysaccharide was dissolved in purified water (5% w / w) and blended with plasticized ethylcellulose dispersion in a ratio of 1: 3 (polymer: polymer w: w) (25% TEC, stirred overnight, 15% w / w polymer content). The mixture was stirred for 6 hours and then cast.

A. 3. Film Characterization The film thickness was measured using a thickness gauge (Minitest 600, Erichsen, Hemer, Germany). All film average thicknesses were in the range of 300-340 μm. Water uptake rate and dry mass loss rate,
(I) Simulated gastric juice (0.1M HCl)
(Ii) Simulated intestinal fluid [phosphate buffer pH 6.8 (USP30) with or without 1% pancreatin or 0.75% rat intestine extract]
(Iii) Culture medium inoculated with stool from healthy subjects (iv) Culture medium inoculated with stool from patients with inflammatory bowel disease (v) Gravimetric method upon exposure to stool-free culture medium for comparison It was measured by.

  1.5 g beef cattle extract, 3 g yeast extract, 5 g tryptone, 2.5 g NaCl, and 0.3 g L-cysteine hydrochloride hydrate in 1 L distilled water (pH 7.0 ± 0.2) The culture medium was prepared by dissolving in ethanol followed by sterilization in an autoclave. Feces from patients with Crohn's disease or ulcerative colitis, as well as feces from healthy subjects, were diluted 1: 200 with cysteine-added Ringer's solution, and 2.5 mL of this suspension was diluted to 100 mL with culture medium. A 1.5 × 5 cm piece of film was placed in a 120 mL glass container filled with 100 mL of pre-heated medium and then shaken horizontally at 37 ° C. (GFL3033, Gesselschaft fur Labtechnik, Burgwedel, Germany). Incubation with stool samples was performed under anaerobic conditions (5% CO2, 10% H2, 85% N2). Samples were withdrawn at predetermined time points, excess water was removed, the film was accurately weighed (wet mass), and dried at 60 ° C. until constant weight (dry mass). The water content (%) and dry film mass (%) at time t were calculated as follows.

A. 4). Bacteriological analysis For bacteriological analysis of stool samples, this was diluted 1:10 with cysteine-added Ringer's solution. Prepare 8 additional 10-fold dilutions with cysteine-added Ringer's solution, and add 0.1 mL of each dilution to non-selective modified Columbia blood agar (to determine total culturable cell count) and McConkey agar (against enteric bacteria) Selective). Columbia blood agar plates were incubated at 37 ° C. for 1 week under anaerobic conditions (5% CO 2, 10% H 2, 85% N 2). The number of colonies was low; the main colonies were subcultured and identified based on phenotypic identification criteria. Twenty-five McConkey agar plates were incubated in air at 37 ° C. for 48 hours. Although the number of colonies was small, they were identified using the API 20E system (BioMerieux, Balme-les-Grottes, France). The count was expressed as log CFU / g (colony forming units per gram) in fresh stool.

  A camera (Unit DS-L2, DS camera Head DS-Fi 1, Nikon, Tokyo, Japan) is provided after gram staining for bacteriological analysis of the microbiota generated when the film is incubated with the stool sample. Micrographs were taken using an Axiostar plus microscope (Carl Zeiss, Jena, Germany). Incubation was performed under anaerobic conditions in a sugar-free culture medium containing a very small amount of polypeptide (thus advantageous for using the polysaccharide under investigation as a substrate).

B. Results and Discussion 1. Film properties within the upper GIT The permeability of a polymer system to a drug is strongly dependent on its water content and dry mass, which determines the density and mobility of the macromolecule. For example, with dry hydroxypropylmethylcellulose (HPMC) matrix tablets, the apparent diffusion coefficient of the drug approaches zero, while with a fully hydrated HPMC gel, a diffusivity comparable to that of an aqueous solution can be achieved. It is. As the water content increases, the mobility of the macromolecules and thus the free volume available for diffusion increases significantly. In some systems, the polymer undergoes a transition from the glass phase to the rubber phase as soon as the critical water content is reached. This causes a significant gradual increase in polymer and drug mobility. Therefore, it is possible to obtain important insights about the mobility of macromolecules and hence the permeability of drugs from the water content of the polymer film coating. FIGS. 1a and 1b show the water uptake rates of thin films composed of various types of polysaccharide: polysaccharide ethylcellulose blends in 0.1N HCl and phosphate buffer pH 6.8, respectively. The presence of ethylcellulose in all films can avoid premature dissolution in the upper GIT. All of the polysaccharides studied are water-soluble and aim to provide drug susceptibility to the surrounding environment of the coating. That is, when it reaches the colon, the polysaccharide is enzymatically degraded and drug release is initiated. The polysaccharide: polysaccharide ethylcellulose blend ratio in FIG. 1 is constant or 1: 3. As is apparent, the water uptake rate and water uptake rate are significantly dependent on the type of polysaccharide. An ideal film coating that allows for colon targeting should be one that takes up very small amounts of water at low rates in both media to prevent premature drug release in the upper GIT. As can be seen, blends of ethylcellulose with BMD or pea starch are the most promising for this purpose. A plasticized ethylcellulose film (open circles) that does not contain a water-soluble polysaccharide takes up only a small amount of water.

  In addition to the water uptake rate, the dry mass loss behavior of the thin polymer film also serves as an indicator for the permeability of the coating to the drug and thus the possibility of suppressing premature release within the upper GIT. If the film loses a significant amount of dry mass upon exposure to the release medium, the coating will be permeable to many drugs, especially those of low molecular weight such as 5-aminosalicylic acid (5-ASA, 153.1 Da). Can be expected. FIGS. 2a and 2b show the thin films composed of various polysaccharide: polysaccharide ethylcellulose blends (constant ratio = 1: 3) upon exposure to 0.1N HCl and phosphate buffer pH 6.8, respectively. It shows the experimentally determined loss of dry mass. The ideal film only loses a small amount of dry mass at a low rate (or no mass at all) and ensures a dense polymer network that is less permeable to the incorporated drug under these conditions To do. As can be seen, the dry mass loss of pea starch-containing and BMD-containing films is very low even after exposure to these release media for up to 8 hours. The observed decrease in dry mass is due, at least in part, to the leaching of the water soluble plasticizer triethyl citrate (TEC, used to plasticize aqueous ethyl cellulose dispersion) into the bulk fluid. It is possible. In addition, some of the water-soluble polysaccharides can leach out of the film. A plasticized ethylcellulose film (white circles) that does not contain a water-soluble polysaccharide loses only a small amount of water, regardless of the type of release medium. However, the permeability of intact ethylcellulose films is known to be very low for many drugs, due at least in part to the low water uptake rate and water uptake rate of these systems. Is possible. For this reason, intact ethylcellulose films are also used as moisture protective coatings. The increase in water uptake rate and water uptake rate of the blend system (FIG. 1) causes a significant increase in polymer chain mobility and thus increased TEC mobility, so the loss of water soluble plasticizer TEC in the bulk fluid. Note that can be expected to be significantly more pronounced in films containing 25% (w / w) water-soluble polysaccharides compared to pure (plasticized) ethylcellulose films.

  It should be pointed out that the results shown in FIG. 2 were obtained in the absence of any enzyme. That pancreatic enzymes can degrade certain polysaccharides, thus inducing significant mass loss and water uptake under in vivo conditions, which can result in increased permeability of the film to the drug Is well known. To clarify the importance of this phenomenon, in the phosphate buffer pH 6.8, in the presence of pancreatin (= mixture containing amylase, protease and lipase) and from the rat intestine (amylase and In the presence of sucrase, isomaltase and glucosidase), the water uptake rate and dry mass loss behavior of the thin film were also measured (FIG. 3). As is apparent, the addition of these enzymes did not significantly affect the resulting water uptake rate and dry mass loss rate of the films studied. Thus, the film does not function as a substrate for these enzymes.

B. 2. Film Properties in the Colon Once the colon is reached, the polymeric film coating must be permeable to the drug. This can be induced, for example, by (partial) enzymatic degradation. Importantly, the concentration of a particular enzyme is much higher in the colon than in the upper GIT. This includes enzymes produced by the natural microflora of the colon (this part of GIT contains much more bacteria than the stomach and small intestine). However, much attention must be paid when using this type of colon targeting technique, as the microbiota of patients suffering from inflammatory bowel disease can be significantly different from that of healthy subjects . Therefore, the drug delivery system must be adapted to the patient's disease state. Table 1 shows the concentration of bacteria determined, for example, in stool samples from healthy subjects, as well as Crohn's disease patients and ulcerative colitis patients incorporated in this study. Importantly, there is a significant difference, especially with respect to the concentration of bifidobacteria (which can degrade complex polysaccharides by multiple extracellular glycosidases) and Escherichia coli, and these bacteria are found in patients with inflammatory bowel disease. Compared with stool, it was present at a considerably higher concentration in stool of healthy subjects. In contrast, stool samples from Crohn's disease patients and ulcerative colitis patients were lactose-negative E. coli, Citrobacter freundii, K. pneumoniae not detected in healthy subjects. (Klebsiella pneumoniae), Klebsiella oxytoca, and Enterobacter cloacae. There are therefore fundamental differences in the quality and quantity of the microflora that must be taken into account. That is, a polymer film coating that allows colon targeting under physiological conditions in healthy volunteers may not work well under pathophysiological conditions in the patient's disease state. To address this very serious problem, which is almost always ignored, stool samples from patients with Crohn's disease and ulcerative colitis, as well as exposure to stool from healthy subjects, and pure cultures for comparison The water uptake and dry mass loss of thin films composed of various types of polysaccharide: polysaccharide ethylcellulose blends upon exposure to the media were examined (FIG. 4). A suitable film must take up a significant amount of water and exhibit significant dry mass loss upon exposure to the patient's feces to induce drug release at sites of inflammation in the colon. As can be seen in FIGS. 4a and 4b, ethyl cellulose: BMD and ethyl cellulose: pea starch (these are the two most promising according to the results described above obtained in the medium simulating the contents of the upper GIT. Films based on polymer blends of the type show significant water uptake and dry mass loss upon exposure to feces of Crohn's disease patients, ulcerative colitis patients and even healthy subjects. Other types of polymer blends are also promising regarding the water uptake and dry mass loss behavior of films exhibited upon exposure to stool samples (and more appropriate than ethylcellulose: BMD blends and ethylcellulose: pea starch blends) Note that it looks like this. However, these systems already take up a significant amount of water and significantly lose dry mass upon contact with the medium simulating the contents of the upper GIT (FIGS. 1 and 2).

  The fact that the polymer film studied serves as a substrate for stool bacteria from patients with inflammatory bowel disease is further confirmed by analysis of the microbiota generated during film exposure to stool samples at 37 ° C under anaerobic conditions (Fig. 5). As is apparent, certain types of bacteria grew when incubated with the blend film. Importantly, this phenomenon is quite beneficial to the GIT ecosystem of diseased patients and can be expected to normalize the microbiota in the colon. This very promising prebiotic effect appears in addition to the drug targeting effect. Biological samples incubated without any polymer film or with pure (plasticized) ethylcellulose film showed significantly less bacterial growth (FIG. 5).

  Novel polymeric film coatings with confirmed colon targeting are water-insoluble polymers: polysaccharide blends, specifically ethyl cellulose: BMD blends, ethyl cellulose: pea starch blends, ethyl cellulose: MD, adapted to the patient's disease state. Blends, ethylcellulose: EURYLON® 6A-PG blend, ethylcellulose: EURYLON® 6HP-PG blend, and ethylcellulose: EURYLON® 7A-PG blend. Importantly, the rate and degree of water uptake and dry mass loss in the medium simulating the contents of the upper GIT is low, and the water uptake and dry weight when in contact with feces from patients with inflammatory bowel disease It is possible to combine increasing losses. Changes in the composition of the microflora in the patient's colon suggest that these polysaccharides may function as substrates for diseased colonic bacteria and likely have beneficial effects on the patient's GIT ecosystem. Is done. Thus, the new knowledge gained provides the basis for the development of new polymeric film coatings that can deliver drugs specifically to the colon. Importantly, these macromolecular barriers are adapted to the pathology of the target site of the disease state.

Example 2
A. Materials and Methods 1. Materials Pregelatinized hydroxypropyl pea starch (PS HP-PG) (LYCOAT® RS780, Roquette Freres), EURYLON® 7A-PG (acetylated and pregelatinized high amylose maize starch (70% amylose)) ( Roquette Freres, Lestrem, France), EURYLON® 6A-PG (acetylated and pregelatinized high amylose maize starch (60% amylose)) (Roquette Freres, Lestrem, France) and EURYLON® 6HP-PG Hydroxypropylated and pregelatinized high amylose maize starch (60% amylose)) (Roquette Freres, Restrem France), aqueous ethylcellulose dispersion (Aquacoat ECD30, FMC Biopolymer, Philadelphia, USA), triethyl citrate (TEC, Morflex, Greensboro, USA).

A. 2. Fabrication of thin polymer films Thin polymer films were prepared by casting blends of various types of polysaccharides and aqueous ethylcellulose dispersions into a Teflon mold followed by drying at 60 ° C for 1 day. Produced. Water-soluble polysaccharides are purified water (15% w / w, hot water in the case of EURYLON® 7 A-PG, EURYLON® 6 A-PG, and EURYLON® 6 HP-PG) And blended with the plasticized aqueous ethylcellulose dispersion at a ratio of 1: 2, 1: 3, 1: 4, 1: 5 (polymer: polymer w: w) as specified (25 based on ethylcellulose content). 0.0, 27.5, or 30.0% w / w TEC, stirred overnight, 15% w / w polymer content). The mixture was stirred for 6 hours and then cast.

A. 3. Film Characterization The film thickness was measured using a thickness gauge (Minitest 600, Erichsen, Hemer, Germany). All film average thicknesses were in the range of 300-340 μm. The water uptake rate and dry mass loss rate of the film were measured gravimetrically upon exposure to 0.1 M HCl and phosphate buffer pH 6.8 (USP30) as follows. That is, a 1.5 × 5 cm piece was placed in a 120 mL plastic container filled with 100 mL of preheated medium, and then horizontally shaken at 37 ° C. (80 rpm, GFL3033, Gesellshaft for Labortechnik, Burgwedel, Germany) . Samples were withdrawn at predetermined time points, excess water was removed, the film was accurately weighed (wet mass), and dried at 60 ° C. until constant weight (dry mass). The water content (%) and dry film mass (%) at time t were calculated as follows.

A. 4). Mechanical properties of thin films The mechanical properties of dry and wet films were investigated using a texture analyzer (TAXT. Plus, Winopal Forschungsbeddorf, Ahnsbeck, Germany) and a piercing test. A film sample was attached to the film holder (n = 6). A puncture probe (spherical end: 5 mm in diameter) was fixed to a load cell (5 kg), and was driven downward toward the center of the hole of the film holder at a crosshead speed of 0.1 mm / s. The load vs. displacement curve was recorded until film failure, and the following mechanical properties were investigated using this.

  Where F is the load required to pierce the film and A is the cross-sectional area of the edge of the film located in the path.

  Where R represents the radius of the film exposed in the cylindrical bore of the holder and D represents the displacement.

  Where AUC is the area under the load vs. displacement curve and V is the volume of the film located in the die cavity of the film holder.

B. Results and Discussion 1. PS HP-PG: ethylcellulose blend FIG. 6 shows the gravimetric method for thin films composed of various types of PS HP-PG: ethylcellulose blends upon exposure to 0.1 M HCl and phosphate buffer pH 6.8, respectively. Shows the water uptake rate and dry mass loss rate determined by. PS HP-PG is pregelatinized modified starch. As with MD, the resulting degree and rate of water penetration into the system increased significantly when the polysaccharide: ethylcellulose ratio was increased from 1: 5 to 1: 2 (FIG. 12, upper side). Figure). Again, this can be due to the higher hydrophilicity of the polysaccharide compared to ethylcellulose. Properly increasing the coating level would probably be necessary to suppress the early release of water soluble low molecular weight drugs in the upper GIT at high initial PS HP-PG content. Also, the rate and degree of dry mass loss of the film increased significantly with increasing PS HP-PG content due to partial TEC and polysaccharide leaching. In either case, the rate and degree of water penetration and dry mass loss is due to the pH-dependent ionization of SDS, as described above, in phosphate buffer pH 6.8 compared to 0.1 M HCl. It was bigger. As with the MD: ethylcellulose blend, the mechanical stability of the PS HP-PG: ethylcellulose film can be effectively adjusted by changing the initial ethylcellulose content. This means that the puncture strength,% elongation at break, and energy at break (Table 2) in the dry state at room temperature, as well as in the wet state upon exposure to 0.1M HCl and phosphate buffer pH 6.8. This was also true for mechanical resistance (FIG. 7). Again, the decrease in break energy with time can be due to partial plasticizer and polysaccharide leaching into the bulk fluid, regardless of the type of release medium.

B. 2. EURYLON® 7A-PG: Ethylcellulose blend Thin composed of 1: 2 to 0: 1 EURYLON® 7A-PG: ethylcellulose blend in 0.1M HCl and phosphate buffer pH 6.8 The water uptake rate and dry mass loss rate of the film are shown in FIG. EURYLON® 7A-PG is an acetylated and pregelatinized high amylose maize starch (70% amylose) (Roquette Freres, Lestrem, France). As can be seen, the same trend was observed for the MD: ethylcellulose blend and the PS HP-PG: ethylcellulose blend. That is, (i) the rate and degree of water uptake increases with decreasing ethylcellulose content, (ii) the rate and degree of dry mass loss increases with increasing polysaccharide content, (iii) These effects were more pronounced in phosphate buffer pH 6.8 than in 0.1M HCl. Importantly, the water content of the film upon exposure to phosphate for 2 hours was fairly large, about 50% w / w. Therefore, a high initial EULYLON® 7A-PG content will likely require a high coating level to suppress the early release of the water soluble low molecular weight drug within the upper GIT. Importantly, when dry at room temperature, the mechanical resistance of the EURYLON® 7A-PG: ethylcellulose-based film is higher than that of films composed of MD: ethylcellulose blends and PS HP-PG: ethylcellulose blends. Significantly higher (Table 2). However, these differences were secondary when the film was exposed to 0.1 M HCl and phosphate buffer pH 6.8, regardless of the type of release medium (FIG. 9). Importantly, again, it was possible to efficiently adjust the mechanical stability of the film by changing the polymer blend ratio.

B. 3. EULYLON® 6A-PG: ethylcellulose blend and EURYLON® 6HP-PG: ethylcellulose blend EURYLON® 6A-PG is an acetylated and pregelatinized high amylose maize starch (60% amylose) (Roquette). EURYLON® 6HP-PG is a hydroxypropylated and pregelatinized high amylose maize starch (60% amylose) (Roquette Freres, Lestrem, France). Interestingly, the dry mass loss of thin films composed of EURYLON® 6A-PG: ethylcellulose blend and EURYLON® 6HP-PG: ethylcellulose blend was 0.1 M HCl and phosphate buffer, respectively. Upon exposure to pH 6.8, it was not as pronounced as the other polymer blends studied (FIGS. 10 and 12, lower diagram). This was true for both the rate and degree of dry mass loss, as well as for all polymer blend ratios studied. In contrast, the rate and degree of water uptake of these films is similar to that of other polysaccharide: ethylcellulose blends upon exposure to various release media, with high initial polysaccharide content. In some cases, a water content of about 50% w / w was reached after 1-2 hours exposure to phosphate buffer pH 6.8 (FIGS. 10 and 12, upper diagram). Therefore, EURYLON® 6A-PG: ethylcellulose blend and EULYLON® 6HP-PG: ethylcellulose blend inhibit early release of water soluble low molecular weight drugs in the upper GIT at low initial ethylcellulose content Therefore, a high coating level will probably be required. As can be seen in Table 2, the mechanical properties of thin films composed of these types of polymer blends dried at room temperature are similar to those of the EURYLON® 7A-PG: ethylcellulose blend with the same blend ratio. It was similar. As with the latter blend, exposure to 0.1 M HCl or phosphate buffer pH 6.8 resulted in mechanical stability of the macromolecular network regardless of the type of release medium and the blend ratio of the polymer. A decrease occurred (Figures 11 and 13). Importantly, the desired system stability can again be effectively adjusted by changing the polymer blend ratio.

  The main of thin polymer films composed of polysaccharide: water insoluble polymer blends that show interesting potential to provide site-specific drug delivery to the colon (and are adapted to the pathophysiology of patients with inflammatory bowel disease) The properties can be effectively tuned by changing the blend ratio of the polymer and the type of polysaccharide. This includes the rate of water uptake before and during exposure to an aqueous medium that simulates the contents of the upper GIT, and the rate of dry mass loss, as well as the mechanical properties of the film. Thus, a wide range of film coating properties can be readily provided, and can be tailored to the respective drug treatment needs (eg, osmotic activity and dosage of the core formulation).

Example 3
A. Materials and Methods 1. Materials Pea starch N-735 (pea starch, Roquette Freres, Restrem, France), Aquacoat ECD30 (aqueous ethyl cellulose dispersion, FMC Biopolymer, Brussels, Belgium, Triethyl citrate (TEC, Morflex A, NC, 5). -Aminosalicylic acid (5-ASA, Sigma-Aldrich, Isled'Abeau Chesnes, France), microcrystalline cellulose (Avicel PH101, FMC Biopolymer), bentonite and polyvinylpyrrolidone (PVP, PovidoneMr.) , France), pancreatin (from mammalian pancreas = mixture of amylase, protease and lipase) and pepsin (Fisher Bioblock, Illkirch, France), extracts from beef cattle and yeast and even tryptone (= pancreatic juice digestion of casein) (Becton, Dickinson and Company, Franklin Lakes, NJ, USA), L-cysteine hydrochloride hydrate (Acros Organics, Geel, Belgium), Cysteine Ringer's solution (Merck, Darmstadt, Germany).

A. 2. Free film preparation A blend of pea starch and aqueous ethylcellulose dispersion (plasticized with 25% TEC) was cast on a Teflon mold and dried under control (60 ° C for 1 day) ) To produce a thin free film. Pea starch was dispersed in purified water at 65-75 ° C. (5% w / w). An aqueous ethylcellulose dispersion (15% w / w solids content) was plasticized with 25% TEC (w / w, based on the solids content of the dispersion) for 24 hours. Pea starch and ethylcellulose dispersions were blended at room temperature in the following ratios: 1: 2, 1: 3, 1: 4, and 1: 5 (polymer: polymer, w: w). The mixture was stirred for 6 hours and then cast.

A. 3. Characteristic analysis of free film The film thickness was measured using a thickness gauge (Minitest 600, Erichsen, Hemer, Germany). All film average thicknesses were in the range of 300-340 μm.

  The rate of water uptake and dry mass of the film upon exposure to (i) simulated gastric fluid (0.1 M HCl) and (ii) simulated intestinal fluid [phosphate buffer pH 6.8 (USP32)] at 37 ° C. as follows: The loss rate was measured by gravimetry. A 1.5 cm × 5 cm piece was placed in a 120 mL plastic container filled with 100 mL of preheated medium, followed by horizontal shaking at 37 ° C. (80 rpm, GFL3033, Gesselschaft for Labortechnik, Burgwedel, Germany). Samples were withdrawn at predetermined time points, excess water was removed, the film was accurately weighed (wet mass), and dried at 60 ° C. until constant weight (dry mass). The water content (%) and dry film mass (%) at time t were calculated as follows.

  Using dry texture and exposure to 0.1M HCl and phosphate buffer pH 6.8 (in wet condition) using a texture analyzer (TAXT. Plus, Winopal Forschungsbeddorf, Ahnsbeck, Germany) and piercing test The physical properties were investigated. A film sample was attached to the film holder (n = 6). A puncture probe (spherical end: 5 mm in diameter) was fixed to a load cell (5 kg), and was driven downward toward the center of the hole of the film holder at a crosshead speed of 0.1 mm / s. The load vs. displacement curve was recorded until film failure, and the following mechanical properties were investigated using this.

  Where F is the load required to pierce the film and A is the cross-sectional area of the edge of the film located in the path.

  Where R represents the radius of the film exposed in the cylindrical bore of the holder and D represents the displacement.

  Where AUC is the area under the load vs. displacement curve and V is the volume of the film located in the die cavity of the film holder.

A. 4). Preparation of coated pellets Drug (5-aminosalicylic acid, 5-ASA) -carrying pellet starter core (diameter: 0.7-1.0 mm, 60% 5-ASA, 32% fine) by extrusion and subsequent spheronization Crystalline cellulose, 4% bentonite, 4% PVP) were prepared as follows. That is, each powder was blended with a high speed granulator (Gral 10, Collette, Antwerp, Belgium) and purified water was added until a uniform mass was obtained (41 g water for 100 g powder blend). The wet mixture was passed through a cylinder extruder (SK M / R, hole: 1 mm diameter, 3 mm thickness, rotation speed: 96 rpm, Alexanderwerk, Remscheid, Germany). Subsequently, the extrudate was spheronized at 520 rpm for 2 minutes (Spheroniser Model 15, Calveva, Dorset, UK) and dried at 40 ° C. for 30 minutes using a fluidized bed (ST15, Aeromatic, Muttenz, Switzerland). A size fraction of 0.7 to 1.0 mm was obtained by sieving. Next, various pea starch: ethylcellulose blends were used with fluid bed coaters (Strea1, Aeromatic-Fielder, Bubendorf, Switzerland) with Wurster inserts at 5, 10, 15 or 20% (w / w). These drug loaded starter cores were coated until weight gain was achieved. A coating formulation was prepared in the same manner as the dispersion used in film casting (as described in Section 2.2 Free film preparation). The process parameters were as follows: Inlet temperature = 39 ± 2 ° C., product temperature = 40 ± 2 ° C., spray rate = 1.5-3 g / min, atomization pressure = 1.2 bar, nozzle diameter = 1.2 mm. The pellet was then fluidized for an additional 10 minutes and subsequently cured in an oven at 60 ° C. for 24 hours.

A. 5. Drug release from coated pellets Drug release from coated pellets was measured in a medium that simulated conditions in the lower gastrointestinal tract.

  Upper gastrointestinal tract: 100 mL lysis medium, ie 0.1 M HCl (optionally containing 0.32% pepsin) for the first 2 hours, followed by phosphate buffer for 9 hours The pellets were placed in a 120 mL plastic container filled with pH 6.8 (USP32) (optionally containing 1% pancreatin). The flask was agitated using a horizontal shaker (80 rpm, GFL 3033). A 3 mL sample was drawn at a given time point and analyzed by UV spectrophotometry for drug content (λ = 302.6 nm in 0.1 M HCl, λ = 330.6 nm in phosphate buffer pH 6.8). (UV-1650, Shimadzu, Champs sur Marne, France). Prior to UV measurement, the sample was centrifuged at 11000 rpm for 15 minutes in the presence of enzyme followed by filtration (0.2 μm). Each experiment was performed in triplicate.

  Whole gastrointestinal tract: USP apparatus 3 (Bio-Dis, Varian, Paris, France) (immersion rate = 10 dpm) was used to expose the pellet to 0.1 M HCl (USP32) for 2 hours, followed by phosphate buffer Exposure to pH 6.8 for 9 hours. Then (i) a 100 mL culture medium inoculated with feces from patients with inflammatory bowel disease, (ii) a culture medium inoculated with bifidobacteria, or (iii) a culture medium without feces and bacteria for comparison. The pellet was transferred to a filled 120 mL flask. Samples were stirred (50 rpm) at 37 ° C. under anaerobic conditions (5% CO 2, 10% H 2, 85% N 2). 1.5 g beef cattle extract, 3 g yeast extract, 5 g tryptone, 2.5 g NaCl, and 0.3 g L-cysteine hydrochloride hydrate in 1 L distilled water (pH 7.0 ± 0.2) The culture medium was prepared by dissolving in ethanol followed by sterilization in an autoclave. Feces from patients suffering from Crohn's disease or ulcerative colitis were diluted 1: 200 with cysteine-added Ringer's solution and 2.5 mL of this suspension was diluted to 100 mL with culture medium. At a given time point, a 2 mL sample was withdrawn, centrifuged at 13000 rpm for 5 minutes, filtered (0.22 μm) and analyzed by HPLC for its drug content (ProStar 230, Varian). The mobile phase consisted of 10% methanol and 90% aqueous acetic acid solution (1% w / v) (Siew et al., 2000a). The sample was injected into a Pursuit C18 column (150 × 4.6 mm, 5 μm). The flow rate was 1.5 mL / min. The drug was detected by UV spectrophotometry at λ = 300 nm.

  Drug release from newly prepared pellets (unless otherwise stated) and further stored in open glass vials for 1 year at room temperature (23 ± 2 ° C) and ambient relative humidity (55 ± 5%) Was measured.

B. Results and Discussion 1. Thin film water uptake and dry mass loss Ideally, polymer film coatings that allow site-specific drug delivery to the colon effectively inhibit drug release in the upper gastrointestinal tract, ie, stomach and small intestine Must be a thing. Thus, the film coating (which surrounds the drug carrier) must be poorly permeable to the drug when exposed to media that simulates the contents of these organs (early drug release and subsequent blood release). To avoid absorption into the flow). If a polymeric film coating incorporates a significant amount of water or loses a significant amount of dry mass upon exposure to a bulk fluid, its permeability to drug molecules can be expected to increase significantly []. To this end, (a) two hours of exposure to 0.1 M HCl (simulating stomach contents) and (b) phosphate buffer pH 6.8 (simulating small intestine contents) During 8 hours exposure to water, the water uptake rate and dry mass loss rate of the thin pea starch: ethylcellulose film were monitored. FIG. 14 shows the experimentally determined water content of the film as a function of time. The pea starch: ethylcellulose blend ratio was varied from 1: 2 to 1: 4 as specified. For comparison, a film (black triangle) composed only of (plasticized) ethylcellulose was also tested. As can be seen, the rate and degree of water uptake increased with increasing pea starch content due to the hydrophilic nature of the polymer. Importantly, water uptake was limited in all cases (less than 30%). A slightly higher water uptake rate and degree at pH 6.8 compared to pH 1.2 (FIG. 14b vs. 14a) is probably present in the aqueous ethylcellulose dispersion used to make the film. This may be due to the presence of sodium dodecyl sulfate (SDS) that functions as a stabilizer. At low pH, SDS is protonated and uncharged, but at pH 6.8 it is negatively charged because it is deprotonated. Therefore, its hydrophilicity is increased and water penetration into the film is promoted [].

  FIG. 15 shows experimentally measured dry mass loss rates of various pea starch: ethylcellulose films upon exposure to (a) 0.1 M HCl and (b) phosphate buffer pH 6.8. . As can be seen, the rate and degree of dry mass loss increased slightly with increasing pea starch content. This is due to the fact that this polysaccharide swells significantly upon contact with water, thereby promoting the leaching of water-soluble film compounds (eg, water-soluble plasticizer TEC []) into the surrounding bulk fluid. is there. Minimal mass loss was observed with pea starch-free films regardless of medium type. This can be due to the fact that ethylcellulose exhibits poor swellability and permeability when in contact with aqueous media. It effectively hinders the leaching of water soluble compounds.

  Thus, the observed rate of water uptake and dry mass loss of pea starch: ethylcellulose films is very promising for the possibility of using this film as a barrier membrane to prevent drug release in the stomach and small intestine. If desired, the film thickness and / or ethyl cellulose content may be increased. However, care must be taken that a sufficient amount of pea starch is present in the coating. Because this compound is intended to induce the onset of drug release in the colon (degraded by enzymes secreted from colonic bacteria).

B. 2. Mechanical properties of thin films In addition to limited water uptake and dry mass loss, polymer film coatings intended for site-specific drug delivery to the colon must provide sufficient mechanical stability. Due to the physiological movements of the stomach and small intestine, mechanical stress is applied to the coated formulation. If the film coating is fragile, it will crack and the drug will be released rapidly through the water-filled channel. A texture analyzer and piercing test were used to evaluate the mechanical stability of the pea starch: ethylcellulose blend studied. FIG. 16 shows (a) the puncture strength at break, (b) the elongation at break%, and (c) the energy required to break the thin polymer film in the dry state. As can be seen, the mechanical stability of the film increased significantly with increasing ethylcellulose content. Interestingly, all values are relatively high, suggesting that film coatings with normal thickness can withstand the mechanical stress experienced in the gastrointestinal tract in vivo.

  However, it should be pointed out that the results shown in FIG. 16 were obtained using a dry film. Upon contact with an aqueous bulk fluid, the mechanical properties of the polymer film coating can change significantly due to, for example, compound leaching into the surrounding bulk fluid and / or plasticizing effects of water. [Erreur! Signon non defini. , Erreur! Signon non defini]. Figures 17a and 17b show the respective energies required to break wet films of various compositions. As is apparent, the mechanical strength of all films decreased with increasing exposure time, regardless of the bulk fluid type. This can be at least partly due to the leaching of the water-soluble plasticizer TEC into the bulk fluid [Erreur! Signon non defini. ]. As expected, the increase in ethylcellulose content caused an increase in the energy required for film breakage in both media. Importantly, observations suggest (at commonly used coating levels) that all film coatings will probably withstand the mechanical stresses that are experienced in vivo in the gastrointestinal tract even when wet.

B. 3. Drug release in the upper gastrointestinal tract Ideally, the drug should be released from the formulation in the stomach and small intestine at all or only slightly. The solid curve in FIG. 18 shows 0, 5, 10, 15, and 20 in 37M C. in 0.1M HCl (2 hours) followed by phosphate buffer pH 6.8 (9 hours). Figure 3 shows experimentally determined drug release rates from pellets coated with pea starch: ethylcellulose 1: 2 at a coating level of% (w / w). As can be seen, 5-aminosalicylic acid was rapidly released from uncoated pellets as well as pellets coated only with 5% pea starch: ethylcellulose 1: 2. This can be due, at least in part, to water uptake and dry mass loss of these film coatings upon exposure to the release medium, along with insufficient thickness of the polymer barrier (FIG. 14). And 15). Importantly, at 10% (w / w) and higher coating levels, drug release was effectively delayed (perhaps due to increased diffusion path length and increased film coating mechanical stability). .

  However, it should be pointed out that the presence of enzymes in the gastrointestinal tract in vivo can significantly affect the properties of the film coating, for example due to partial polymer degradation. For this purpose, (i) 0.1M HCl containing 0.32% pepsin (2 hours) followed by (ii) phosphate buffer pH 6.8 (9 hours) containing 1% pancreatin The drug release from the attached pellet was measured. Each result is shown as a dotted curve in FIG. As is apparent, in all cases, the drug release rate increased only slightly. Therefore, the importance of such enzymatic degradation in vivo is probably limited.

  Increasing the relative ethylcellulose content of the film resulted in a decrease in the rate and degree of water uptake and dry mass loss (FIGS. 14 and 15) as well as an increase in the mechanical stability of the dry and wet films (FIGS. 16 and 15). 17), so drug release from pellets coated with pea starch: ethylcellulose 1: 3, 1: 4, and 1: 5 blends at various coating levels was measured (FIG. 19). Interestingly, in these cases, even 5% (w / w) film coating can reduce the rate of drug release. However, a coating level of 10% or more is more appropriate because drug release is almost completely suppressed within the observation period.

  Based on the results obtained (FIGS. 14-19), pellets coated with pea starch: ethylcellulose 1: 4 at 15% and 20% coating levels were selected for further testing.

B. 4). Drug release in the entire gastrointestinal tract When the formulation reaches the colon, the film coating must be permeable to the drug and release the drug in a time-controlled manner. FIG. 20a shows (i) 0.1 M HCl for 2 hours, followed by (ii) 9 hours, phosphate buffer pH 6.8, and (c) 10 hours for patients with inflammatory bowel disease Of 5-aminosalicylic acid from pellets coated with pea starch: ethylcellulose 1: 4 at a coating level of 15% and 20% (w / w) in culture medium inoculated with fecal samples from Shows the measured release (solid curve). For comparison, drug release upon exposure to culture media without feces is also shown (dotted curve). As can be seen, drug release was initiated as soon as the pellet contacted the stool sample. This may be due to (at least partial) degradation of pea starch by enzymes secreted by bacteria present in the patient's colon. The reduction in polymer molecular weight and subsequent diffusion of degradation products into the surrounding bulk fluid makes the remaining macromolecular network more mobile. Thus, the mobility of drug molecules within the film coating is also increased, thereby increasing the release rate. In contrast, the drug release rate remained low upon exposure to culture medium without feces (dotted curve in FIG. 20a). This confirms that drug release is triggered by an enzyme present in the colon of an IBD patient. From a practical point of view, a 15% coating level seems to be preferred over a 20% coating level (which may result in drug release that is too low in the colon).

  Because it is difficult to ensure a regular supply of new stool samples from patients with inflammatory bowel disease (and samples cannot be snap frozen or lyophilized without significant damage to the microflora) It would be highly desirable to provide an alternative type of release medium that simulates a condition in the patient's colon. In drug delivery systems that are sensitive to the presence of bacterial enzymes, care must be taken that the bulk fluid contains a critical type and amount of bacteria. In this test, culture medium inoculated with bifidobacteria was tested as a powerful alternative to culture medium inoculated with fresh stool samples. FIG. 20b is shown in FIG. 20a upon exposure to culture medium (10 hours) inoculated with 0.1 M HCl (2 hours), phosphate buffer pH 6.8 (9 hours), and bifidobacteria. Shows the observed drug release rate from the same type of pellet. Comparing FIGS. 20a and 20b, the culture medium inoculated with bifidobacteria is promising as a substitute for new stool samples from IBD patients, especially in routine applications (eg quality control in large-scale production). It becomes clear to show.

  Similar results were obtained using dibutyl sebacate as the plasticizer (data not shown), confirming that the efficiency of pea starch in controlled release delivery is not plasticizer dependent.

B. 5. Storage stability A very important aspect from a practical point of view is the long-term stability of the controlled drug delivery system. The formulation should ideally be stable for at least 3 years. In the case of a polymer coated delivery system, the resulting drug release rate can eventually increase with increasing shelf life due to, for example, drug migration into the film coating. FIG. 21 shows coating with pea starch: ethylcellulose 1: 4 before, after 1 year storage in open glass vials (solid and dotted curve) at coating levels of 10, 15 and 15% (as specified) The drug release rate from the prepared pellet is shown. The system was exposed to 0.1 M HCl for 2 hours followed by 9 hours exposure to phosphate buffer pH 6.8. As can be seen, the release rate was held unchanged during long-term storage. The same is true for pellets coated with pea starch: ethylcellulose 1: 2, 1: 3, and 1: 5 at coating levels of 10, 15, and 20% (data not shown). Thus, the proposed drug delivery system is stable for a long time.

C. CONCLUSION Pea starch: ethylcellulose-based film coatings have been proposed as potentially promising for site-specific drug delivery to the colon. Drug release from coated pellets can be effectively inhibited in a medium that simulates the contents of the stomach and small intestine. However, as soon as the device contacts the stool sample, drug release is initiated due to partial degradation of pea starch by enzymes secreted from bacteria present in the colon of patients with inflammatory bowel disease, and Time controlled. Thus, this type of advanced delivery system can avoid early drug release in the upper gastrointestinal tract (and subsequent absorption into the bloodstream) while ensuring drug release at the site of action. To. Thus, undesirable side effects in the rest of the human body can be expected to be minimized, while the therapeutic effect of the drug will likely be optimized.

Example 4
A. Materials and Methods 1 Materials 2,4,6-Trinitrobenzenesulfonic acid (TNBS) (Sigma-Aldrich, Isled'Abeau Chesnes, France), Cysteine Ringer's solution (Merck, Darmstadt, Germany), BMD (NUTRIOSE® FB06te, Roqte) Freres, Restrem, France), pea starch N-735 (pea starch, Roquette Freres, Restrem, France), aqueous ethylcellulose dispersion (Aquacoat ECD30, FMC Biopolymer, Philadelphia, USA, triC) USA), 5-aminosalicylic acid (5-ASA) Sigma-Aldrich, Isled'Abeau Chesnes, France), microcrystalline cellulose (Avicel PH101, FMC Biopolymer, Brussels, Belgium), Polyvinylpyrrolidone (PVP, PovidoneMen), Cooperation Pharm (Trademark) (coated pellet, Ferring, batch number: JX155), Asacol® (coated granule, Meduna, batch number: TX143).

A. 2 Production of pellets with BMD: ethylcellulose coating and pea starch: pellets with ethylcellulose coating 5-aminosalicylic acid (5-ASA) -carrying pellet starter core (diameter: 0.7-1.0 mm) by extrusion and subsequent spheronization , 60% 5-ASA, 32% microcrystalline cellulose, 4% bentonite, 4% PVP). That is, each powder was blended with a high speed granulator (Gral 10, Collette, Antwerp, Belgium) and purified water was added until a uniform mass was obtained (41 g water for 100 g powder blend). The wet mixture was passed through a cylinder extruder (SK M / R, hole: 1 mm diameter, 3 mm thickness, rotation speed: 96 rpm, Alexanderwerk, Remscheid, Germany). Subsequently, the extrudate was spheronized at 520 rpm for 2 minutes (Spheroniser Model 15, Calveva, Dorset, UK) and dried at 40 ° C. for 30 minutes using a fluidized bed (ST15, Aeromatic, Muttenz, Switzerland). A size fraction of 0.7 to 1.0 mm was obtained by sieving.

  Subsequently, 15% (w / w) (BMD: pellets with EC coating) or 20% (w / w) using a fluid bed coater with a Wurster insert (Strea 1, Aeromatic-Fielder, Bubendorf, Switzerland) The resulting drug-loaded starter core is blended with BMD: ethylcellulose 1: 4 blend (BMD: pellet with EC coating) or pea starch: ethylcellulose 1: until a weight gain of (pea starch: pellet with EC coating) is achieved. Coated with 2 blends (pea starch: pellets with EC coating).

  BMD is dissolved in purified water (5% w / w) and blended with the plasticized aqueous ethylcellulose dispersion at a 1: 4 ratio (w / w, based on dry weight of unplasticized polymer) (25% TEC Stir overnight, 15% w / w polymer content) and stir for 6 hours before coating. The drug-loaded pellet core was coated using a fluid bed coater with a Wurster insert (Strea1, Aeromatic-Fielder, Bubendorf, Switzerland) until a weight gain of 15% (w / w) was achieved. The process parameters were as follows: Inlet temperature = 39 ± 2 ° C., product temperature = 40 ± 2 ° C., spray rate = 1.5-3 g / min, atomization pressure = 1.2 bar, nozzle diameter = 1.2 mm. After coating, the beads were fluidized for an additional 10 minutes, followed by curing in a 60 ° C. oven for 24 hours.

  Pea starch was dispersed in purified water at 65-75 ° C. (5% w / w). An aqueous ethylcellulose dispersion (15% w / w solids content) was plasticized with 25% TEC (w / w, based on the solids content of the dispersion) for 24 hours. Pea starch and ethylcellulose dispersion were blended at room temperature in the following ratio: 1: 2 (polymer: polymer, w: w). The mixture was stirred for 6 hours before coating. The drug-loaded pellet core was coated using a fluid bed coater with a Wurster insert (Strea1, Aeromatic-Fielder, Bubendorf, Switzerland) until a 20% (w / w) weight gain was achieved. The process parameters were as follows: Inlet temperature = 39 ± 2 ° C., product temperature = 40 ± 2 ° C., spray rate = 1.5-3 g / min, atomization pressure = 1.2 bar, nozzle diameter = 1.2 mm. The pellet was then fluidized for an additional 10 minutes and subsequently cured in an oven at 60 ° C. for 24 hours.

A. 3 Induction of colitis and study design Male Wistar rats (250 g) were used for the in vivo study. This test was conducted in an accredited facility (A35009) of the Institute Pasteur de Lille in accordance with government guidelines (86/609 / CEE). Four animals were bred per cage. All rats had free access to tap water.

  At the start of the experiment (day 0), the rats were divided into 6 groups (5-8 animals / group). Two groups received standard diet (negative and positive control group). Other groups are Pentasa (R) pellets (n = 8), Asacol (R) pellets (n = 8), BMD: pellets with ethyl cellulose coating (n = 8), or pea starch: ethyl cellulose coating A food containing any of the attached pellets (n = 8) was ingested. These four different feeds were prepared using the “food blending” technique. All systems were added to give a dose of 150 mg / kg / day of 5-ASA.

  On day 3, colitis was induced as follows: rats were anesthetized with pentobarbital (40 mg / kg) for 90-120 minutes and a 1: 1 mixture of aqueous 0.9% NaCl solution and 100% ethanol. Intrarectal administration of TNBS (250 μl, 20 mg / rat) dissolved in the solution was performed. Control rats (negative control) received intrarectal administration of vehicle alone (1: 1 mixture of aqueous 0.9% NaCl solution and 100% ethanol). Three days after intrarectal TNBS administration or intrarectal vehicle administration (day 6), the animals were sacrificed.

A. 4 Macroscopic and histological evaluation of colitis Macroscopic and histological indicators of colitis were blindly evaluated by two investigators. A colon specimen located exactly 4 cm above the anal canal was used for histological evaluation according to the Ameho criteria. This grading based on a scale of 0-6 takes into account the degree of inflammatory infiltration, the presence of wrinkles, ulcers or necrosis, and the depth and surface area of the lesion.

A. 5 Statistics All comparisons were analyzed using a non-parametric test (Mann-Whitney test). If the P value was <0.05, the difference was judged to be statistically significant.

B Results and Discussion TNBS-induced colitis is improved by treatment with BMD: ethylcellulose coated pellets and pea starch: ethylcellulose coated pellets.

  The progression of colitis was characterized in animals receiving intrarectal TNBS administration. Control rats (negative control group) sacrificed 3 days after rectal administration of vehicle alone (1: 1 mixture of aqueous 0.9% NaCl solution and 100% ethanol) have macroscopic lesions in the colon. (FIG. 22A). In contrast, severe colitis was induced as early as 3 days after TNBS administration (FIG. 22B). Regarding the histological level, no abnormality was detected in the control rats (FIG. 24: control = negative control group). In contrast, 3 days after TNBS administration, colon histology was characterized by a large ulcer area with neutrophil infiltration, with necrosis spreading deep into the muscle layer (FIG. 24: TNBS). The colon of animals treated with BMD: ethylcellulose coated pellets showed a significant reduction in lesions (FIG. 22). This result was similar to that of rats treated with pea starch: ethylcellulose coated pellets (data not shown). In addition, the effect of treatment with Pentasa® pellets, Asacol® pellets, BMD: ethylcellulose coated pellets, and pea starch: ethylcellulose coated pellets on TNBS-induced colonic lesions was measured by the Ameho score. (FIG. 29). Untreated rats with colitis were examined for comparison. The optimal effect was obtained with BMD: ethylcellulose coated pellets and with pea starch: ethylcellulose coated pellets. Three days after induction of colitis, macroscopic lesions were observed in rats given prophylactic administration of BMD: ethylcellulose-coated pellets and pea starch: ethylcellulose-coated pellets compared to untreated rats with colitis. A significant decrease in score was observed. In response to macroscopic inflammation, histological analysis also revealed that (i) TNBS in the rectum, (ii) TNBS in the rectum and orally Pentasa pellets, (iii) TNBS in the rectum and Orally Asacol® pellets, (v) TNBS in the rectum and orally BMD: ethylcellulose coated pellets, and (v) TNBS in the rectum and orally pea starch: ethylcellulose coated pellets Major differences were confirmed between animals treated with (Figure 24). This was reflected in a significant decrease in the Ameho inflammation score 3 days after TNBS administration (FIG. 23). As is apparent, the administration of BMD: ethylcellulose coated pellets and pea starch: ethylcellulose coated pellets is inflammatory composed of smaller polymorphic inflammatory infiltrates, localized edema, and small focal necrotic lesions. Lesions were reduced (Figure 24). When treated with TNBS, TNBS and Pentasa® pellets, and TNBS and Asacol® pellets, with marked inflammatory infiltration in the lamina propria and necrosis extending deep into the muscle and serosa layers A thickening of the colon wall is evident.

  These results clearly demonstrate the effectiveness of the novel film coating proposed for colon targeting in vivo.

Claims (13)

A controlled release delivery formulation for controlled release of an active ingredient, comprising:
The formulation comprises a core and a coating mixture;
The coating mixture is
o is a polymer mixture of a starch composition comprising a granular pea starch down with at least water-insoluble polymer and o 25 to 45% of amylose content,
The active ingredient is dispersed or dissolved in the core and / or in the coating mixture;
Controlled release delivery formulation. Before SL active ingredient is dispersed or dissolved in said core, controlled release delivery formulation according to claim 1.   3. The controlled release delivery formulation according to claim 1 or 2, wherein the starch composition: water insoluble polymer ratio is from 1: 2 to 1: 8. The controlled release delivery formulation according to any one of claims 1 to 3 , wherein the granular pea starch is a modified granular pea starch, and the modified granular pea starch is stabilized. The controlled release delivery formulation according to any one of claims 1 to 4 , wherein the core has a coating level of 5% to 30%. 6. The controlled release delivery formulation according to claim 5 , wherein the core has a coating level of 10% to 20%. The polymer mixture comprises a soluble plasticizer, controlled-release delivery formulation according to any one of claims 1-6. 8. The controlled release delivery formulation according to claim 7, wherein the polymer mixture comprises a plasticizer at a content of 25-30% w / w based on the water-insoluble polymer content. The water-insoluble polymer is acrylic acid and / or methacrylic acid ester polymer, acrylate or methacrylate polymer or copolymer, polyvinyl ester, polyvinyl acetate, polyacrylic acid ester, and butadiene styrene copolymer, methacrylic acid ester copolymer, ethyl cellulose, cellulose acetate phthalate Controlled release properties according to any one of claims 1 to 8 , selected from the group consisting of: polyvinyl acetate phthalate, shellac, methacrylic acid copolymer, cellulose acetate trimellitate, hydroxypropylmethylcellulose phthalate, zein, starch acetate. Delivery formulation. The plasticizer is Ru der soluble plasticizers, controlled release delivery formulation according to any one of 請 Motomeko 7-9. 11. The controlled release delivery formulation according to claim 10, wherein the plasticizer is selected from the group consisting of polyol, organic ester, oil or glyceride, soy lecithin, alone or as a mixture with one another.   The controlled release delivery formulation according to any one of claims 1 to 11, wherein the controlled release delivery formulation is a multiparticulate formulation. 13. A method for producing a controlled release delivery formulation for controlled release of an active ingredient in the colon of a patient having an imbalance of intracolonic microbiota or in the colon of a healthy subject according to any one of claims 1-12. Because
o The following substances:
- forming a polymer mixture of a starch composition comprising a granular pea starch down with at least one water-insoluble polymer and, 25% to 45% of amylose content,
o coating the active ingredient with the polymer mixture;
Including a method.